Histomorphometric and immunohistochemical analysis of infectious agents, T-cell subpopulations and inflammatory adhesion molecules in placentas from HIV-seropositive pregnant women.

BACKGROUND: The aim of this study was to compare histomorphometric changes and the results of immunohistochemical tests for VCAM, ICAM-1, CD4 and CD8 in normal placentas from HIV-seropositive pregnant women. METHODS: Samples of normal placentas were divided into 2 groups: healthy HIV-seronegative pregnant women (control group = C = 60) and HIV-seropositive women (experimental group = E = 57). Conventional histological sections were submitted to morphometric analysis and evaluated in terms of the immunohistochemical expression of ICAM-1, VCAM, CD4 and CD8. RESULTS: The villi in group E were smaller than those in group C. The median for the CD8+ T cell count was higher in group E than in group C (p = 0.03). Immunohistochemical expression of ICAM-1 was observed in 57% of the cases in group E, compared with 21% of those in group C (p = 0.001). There was no difference in VCAM expression or CD4+ cell counts between groups and no correlation between the data for antiretroviral therapy and morphometric or immunohistochemical data. CONCLUSIONS: The morphometric data showed that placentas of HIV-seropositive pregnant women tend to have smaller villi than those of seronegative women. In addition, immunohistochemical testing for infectious agents helped to identify cases that were positive for microorganisms (6/112) that routine pathological examination had failed to detect. The anti-p24 antibody had a limited ability to detect HIV viral protein in this study (2/57). Correlation of immunohistochemical expression of CD8+ T cells and ICAM-1 with the presence of HIV in the placenta revealed that those expressions can act as biomarkers of inflammatory changes. There was no correlation between the data for antiretroviral therapy and morphometric or immunohistochemical data.

Respiratory viruses in pediatric necropsies: an immunohistochemical study.

Infections of the respiratory system are responsible for the majority of hospitalizations and deaths in pediatric patients in developing countries. We selected 177 necropsies of pediatric patients who died as a result of serious respiratory infections. The histopathological findings and epidemiological data were reviewed, and lung tissue samples were separated for immunohistochemistry testing. Conventional immunohistochemistry techniques were used to detect viral antigens in formalin-fixed, paraffin-embedded (FF-PE) lung tissue samples using a pool of monoclonal antibodies against respiratory viruses (respiratory syncytial virus, influenza A and B, adenovirus, and parainfluenza 1, 2, and 3 viruses) as primary antibodies. The histopathological findings were classified into bronchopneumonia (BCP) and interstitial pneumonitis (IP) patterns. The immunohistochemistry results were compared with histopathological patterns and epidemiological data. Positive results for viruses were found in 34% and 62.5% of the BCP and IP cases, respectively. Males and infants below 1 year of age were more frequent in the group that had positive results for viruses. Acute enteritis was the main cause of hospitalization and sepsis the most frequent cause of death in this group. A clear seasonal distribution was observed, with the majority of cases occurring in the 2nd and 3rd trimesters (autumn and winter) of each year in the period studied. Immunohistochemistry is an affordable and easy-to-perform method for viral-antigen detection in FF-PE tissue samples. Although BCP is a classic histopathological pattern found in bacterial infections, it is possible that children with serious respiratory infections had concomitant viral and bacterial infections, regardless of their previous immunologic state.

Gasometria arterial e inflamação pulmonar de ratos com diferentes tempos de sepse abdominal / Arterial gasometry and lung inflammation in rats with different times of abdominal sepsis

RACIONAL: Sepse é a principal causa de morbi-mortalidade nas vítimas de trauma e em pacientes cirúrgico e apesar de toda tecnologia e terapêutica disponível não há diminuição nestas estatísticas. OBJETIVOS: Avaliar as repercussões ácido-básicas e o grau de injúria pulmonar decorrentes de sepse abdominal em ratos após seis e 24 horas de peritonite fecal através da ligadura e punção do ceco. MÉTODOS: Foram utilizados 40 ratos Wistar, machos, adultos. A amostra foi dividida aleatoriamente em quatro grupos: grupo A (sham/6 h - n=5) submetidos à laparotomia mediana infra-umbilical sem nenhuma outra intervenção; grupo B (sham/24h - n=5) submetidos à laparotomia mediana infra-umbilical sem nenhuma outra intervenção; grupo C (LPC/6 h - n=15) submetidos à ligadura e punção do ceco e grupo D (LPC/24h - n=15) submetidos à ligadura e punção do ceco. Após seis ou 24 horas, conforme o grupo em estudo, os animais foram novamente anestesiados e submetidos as seguintes análises: observação clínica de sinais de sepse, laparotomia através do mesmo acesso anterior e realização de cultura do líquido peritoneal e punção cardíaca para obtenção de amostra sangüínea suficiente para gasometria arterial, hematócrito e leucometria. Procedeu-se eutanásia e os pulmões retirados para análise de edema pulmonar e o infiltrado inflamatório. RESULTADOS: Ocorreram quatro óbitos no grupo D. Verificou-se, em todos os grupos, a presença de acidose mista. Comparando os grupos experimento 6h versus controle 6h foram encontradas duas variáveis significativas - HCO3 (p=0,0015) e BE (p=0,0015) -, demonstrando acidose metabólica mais grave no grupo experimento. Nos grupos controle também se confirmou acidose mista, devido às alterações das variáveis HCO3 (p=0,0079), PO2 (p=0,0079) e SO2 (p=0,0079). A correlação entre o grau de comprometimento pulmonar e o estado metabólico confirma a existência de resposta inflamatória sistêmica evidenciada por aumento de neutrófilos e hemorragia alveolar difusa nos pulmões do rato séptico. CONCLUSÃO: O modelo de LPC foi método eficaz para indução de sepse. A laparotomia e a manipulação de alças intestinais são fatores desencadeantes de acidose mista em ratos, comprovado pelos resultados da gasometria, que demonstrou ser método confiável na detecção de alterações no metabolismo ácido-básico dos ratos estudados.

BACKGROUND: Sepsis is the major cause of morbidity and mortality in trauma victims and surgical patients, and despite of all the technology and therapy available, the statistics for these cases is not diminishing. AIM: To evaluate the acid-basic repercussions and degree of pulmonary injury due to abdominal sepsis in rats after 6 and 24 hours of fecal peritonitis through cecal ligation and puncture. METHODS: CLP in experimental groups. Forty Wistar adult rats were distributed into four groups: A (sham/6 h - n=5) submitted to infra-umbilical median laparotomy, with no other interventions; B (sham/24 h - n=5) submitted to infra-umbilical median laparotomy, with no other interventions; C (CLP/6 h - n=15) submitted to ligation and cecal puncture and D (CLP/24 h - n=15) submitted to ligation and cecal puncture. According to the study group, after a period of six or 24 hours, the animals were once again anesthetized and submitted to the following analysis: clinical observations for sings of sepsis, laparotomy through the same access point made earlier, cultures of peritoneal fluid, cardiac puncture for obtaining sufficient blood samples for arterial gasometry, hematocryte e leucometry analysis. Euthanasia was performed and lungs removed were analyzed for pulmonary edema and inflammatory infiltrate. RESULTS: Four deaths occurred in group D. The presence of mixed acidosis in all groups was verified. When comparing groups B and D, two significant variables were found: HCO3 (p=0,0015) and BE (p=0,0015), demonstrating a more severe metabolic acidosis in the experimental group. Mixed acidosis was also confirmed in the control group, due to alterations regarding HCO3 (p=0,0079), PO2 (p=0,0079) and SO2 (p=0,0079). Correlation between pulmonary commitment and metabolic state confirms the existence of systemic inflammatory response, demonstrated through the increase of neutrophils and diffuse alveolar hemorrhage of the lungs of septic rats. CONCLUSION: The present study demonstrated that CLP was an efficient method to induce sepsis in rats. It was observed that laparotomy and bows' manipulation resulted in mixed acidosis in rats, being verified by gasometrical findings, demonstrating to be a reliable method for the detection of acid-basic metabolic alterations in the rats studied.